Abstract
Introduction
The goal of regenerative endodontic procedures is to preserve and stimulate stem cells
from the apical papilla (SCAPs) to develop the pulp-dentin complex using various growth
factors and scaffolds. We hypothesized that the treatment of SCAPs with vascular endothelial
growth factor (VEGF) or nerve growth factor (NGF) may impact the expression of osteogenic
and dentinogenic markers.
Methods
The optimum concentration of VEGF and NGF on SCAP viability was assessed and introduced
to SCAPs for 6–24 hours. SCAPs were also challenged with Escherichia coli lipopolysaccharide (LPS). Messenger RNA (mRNA) expression of DSPP, DMP1, TGFB1, OCN, SP7, and TWIST1 was examined via quantitative reverse transcription polymerase chain reaction. Immunohistochemistry
was used to verify protein expression. In addition, total RNA from NGF-treated SCAPs
in the presence or absence of LPS was extracted for RNA sequencing.
Results
Compared with untreated cells, NGF-treated SCAPs showed markedly higher levels of
DSPP, DMP1, and TGFB1 mRNAs (>9-fold change, P < .05), and SCAPs treated with both VEGF and NGF showed a significant increase of
DSPP and TGFB1 mRNAs (P < .05). In addition, in LPS-challenged SCAPs, treatment with these growth factors
also exhibited increased expression of DSPP, DMP1, and TGFB1 mRNAs, with the most significant change induced by VEGF (P < .05). Immunohistochemistry confirmed increased dentin sialophosphoprotein, dentin
matrix acidic phosphoprotein 1, and transforming growth factor beta 1 protein expression
in treated SCAPs. RNA sequencing revealed multiple pathways regulated by NGF, including
TGF-β and neurogenic pathways.
Conclusions
VEGF- and NGF-induced dentinogenic/neuronal/healing marker expression in SCAPs indicates
the potential value of applying these growth factors in regenerative endodontic procedures.
Key Words
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Article info
Publication history
Published online: February 26, 2021
Accepted:
February 18,
2021
Received in revised form:
February 9,
2021
Received:
October 27,
2020
Identification
Copyright
© 2021 American Association of Endodontists.
